CN104437280B - A kind of preparation method of hybrid magnetic Nano flower - Google Patents

Abstract

The invention relates to a method for preparing organic/inorganic hybrid magnetic nanoflowers, which is characterized in that: magnetic nano-scale or submicron-scale particles functionalized with surface amine groups or carboxyl groups are used as magnetic sources, and biomacromolecules (such as proteins) , enzymes, etc.) and zinc phosphate (Zn ₃ (PO ₄ ) ₂ ) formed organic/inorganic hybrid nanostructures as the main skeleton, the composite of magnetic particles and organic/inorganic hybrid nanoflowers and the combination of biological macromolecules and Zn ₃ (PO 4 ) 2 ₄ ) The hybrid combination of ₂ utilizes the coordination interaction between Zn ²⁺ and the amine or carboxyl groups on the surface of magnetic particles or biomacromolecules. The prepared organic/inorganic hybrid magnetic nanoflowers have high application value in the fields of adsorption separation and catalytic synthesis according to the difference of biological macromolecules. At the same time, due to the introduction of magnetic components, the materials can be quickly recovered and reused. .

Description

A kind of preparation method of organic/inorganic hybrid magnetic nano flower

technical field

The present invention relates to a method for preparing organic/inorganic hybrid magnetic nanoflowers, in particular to biomacromolecules (proteins, enzymes, etc.), magnetic nanometer or submicron particles, zinc phosphate (Zn ₃ (PO ₄ ) ₂ ) A simple one-step synthesis of three materials to prepare organic/inorganic hybrid magnetic nanoflowers.

Background technique

Organic/inorganic hybrid magnetic composite microspheres integrate organic matter and magnetic inorganic particles, so they can be quickly separated and enriched under a magnetic field, and at the same time, they are easy to modify the surface for functionalization, so they are widely used in biochemistry, medicine, environment, detection , sensing and other fields have extensive and important applications. Among them, the magnetic inorganic particles are generally mainly composed of ferric oxide (Fe ₃ O ₄ ) and ferric oxide (Fe ₂ O ₃ ), and the organic components can be various high molecular polymers (ZL201010202232.7; ZL200610042745.X) , natural polymers (CN201110324675.8; CN201110141162.3), etc. In order to improve the performance, magnetic composite microspheres with high specific surface area have become a research hotspot, but most of them are coated with inorganic mesoporous layers on the surface of magnetic inorganic particles or embedded in the framework of porous polymer microspheres (ZL200910021833.5 ), CN201010120183.2 reported a water-soluble rock-salt type CoO nano-flower-shaped magnetic particle, but no organic/inorganic hybrid magnetic composite microspheres similar to inorganic nano-flower-shaped materials have been reported.

Contents of the invention

technical problem to be solved

In order to avoid the deficiencies of the prior art, the present invention proposes a method for preparing organic/inorganic hybrid magnetic nanoflowers, using biomacromolecules (proteins, enzymes, etc.), magnetic nanoscale or submicron particles, Zn ₃ ( Organic/inorganic hybrid magnetic nanoflowers were synthesized by PO ₄ ) ₂ in a simple one-step method.

Technical solutions

A method for preparing organic/inorganic hybrid magnetic nanoflowers, characterized in that the steps are as follows:

Step 1: uniformly disperse the magnetic particles in water, prepare an aqueous dispersion of magnetic particles with a mass fraction of 0.05-0.15%, and add it to a container with mechanical stirring;

Step 2: adding a soluble zinc salt solution with a concentration of 0.2-0.5 mol/L, and stirring for 0.5-3 hours; wherein the volume ratio of the soluble zinc salt solution to the magnetic particle aqueous dispersion is 1:10-25;

Step 3: adding 0.2-1 g/L phosphate buffer solution, wherein the volume ratio of phosphate buffer solution to soluble zinc salt solution is 10-25:1; the pH range of the phosphate buffer solution is 7.2-7.4;

Step 4: Under mechanical stirring, after reacting at room temperature for 2-8 hours, magnetic field separation, water washing, and freeze-drying are performed to obtain organic/inorganic hybrid magnetic nanoflowers.

The soluble zinc salt is: one or more of zinc acetate and its hydrate, zinc nitrate and its hydrate, zinc chloride and its hydrate, or zinc sulfate and its hydrate in any ratio.

The soluble biomacromolecule is bovine serum albumin, pepsin, trypsin, lipase, amylase or papain.

The phosphate is: sodium dihydrogen phosphate, disodium hydrogen phosphate and water or potassium dihydrogen phosphate, dipotassium hydrogen phosphate and water or sodium dihydrogen phosphate, dipotassium hydrogen phosphate and water or potassium dihydrogen phosphate, dihydrogen phosphate sodium and water.

Beneficial effect

A method for preparing organic/inorganic hybrid magnetic nanoflowers proposed by the present invention uses magnetic nanoscale or submicron-scale particles functionalized with surface amine or carboxyl groups as the magnetic source, and biomacromolecules (such as proteins, enzymes, etc.) The organic/inorganic hybrid nanostructure formed with zinc phosphate (Zn ₃ (PO ₄ ) ₂ ) is used as the main skeleton, the composite of magnetic particles and organic/inorganic hybrid nanoflowers and the combination of biological macromolecules and Zn ₃ (PO ₄ ) ₂ Hybrid recombination utilizes the coordination interaction between Zn ²⁺ and the amine or carboxyl groups on the surface of magnetic particles or biomacromolecules. The prepared organic/inorganic hybrid magnetic nanoflowers have high application value in the fields of adsorption separation and catalytic synthesis according to the difference of biological macromolecules. At the same time, due to the introduction of magnetic components, the materials can be quickly recovered and reused. .

Description of drawings

Figure 1 is a flow chart of the preparation process of organic/inorganic hybrid magnetic nanoflowers

Figure 2 is the SEM photo of bovine serum albumin/Zn ₃ (PO ₄ ) ₂ /Fe ₃ O ₄ hybrid magnetic nanoflowers

detailed description

Now in conjunction with embodiment, accompanying drawing, the present invention will be further described:

Example 1: Preparation of bovine serum albumin/Zn ₃ (PO ₄ ) ₂ /Fe ₃ O ₄ hybrid magnetic nanoflowers

Disperse Fe ₃ O ₄ particles evenly in water to obtain a water dispersion of Fe ₃ O ₄ particles with a mass fraction of 0.05-0.15%, and add it to a three-necked flask with mechanical stirring; prepare acetic acid with a concentration of 0.2-0.5mol/L Add the zinc aqueous solution into the three-necked flask, and stir it mechanically for 0.5-3h. Then add _0.2-1g /L bovine serum albumin solution, the solvent is phosphate buffer solution, wherein the volume ratio of _Fe3O4 particle water dispersion, bovine serum albumin solution and zinc acetate aqueous solution is 10-25: 10～25:1. Under mechanical stirring, after reacting at room temperature for 2-8 hours, the bovine serum albumin/Zn ₃ (PO ₄ ) ₂ /Fe ₃ O ₄ hybrid magnetic nanoflowers are obtained through magnetic field separation, water washing and freeze-drying.

Example 2: Preparation of bovine serum albumin/Zn ₃ (PO ₄ ) ₂ /Fe ₂ O ₃ hybrid magnetic nanoflowers

Disperse Fe ₂ O ₃ particles evenly in water to obtain a water dispersion of Fe ₂ O ₃ particles with a mass fraction of 0.05-0.15%, and add it to a three-necked flask with mechanical stirring; prepare chlorine with a concentration of 0.2-0.5mol/L Zinc chloride aqueous solution was added into a three-necked flask, and mechanically stirred for 0.5-3 hours. Then add 0.2-1g/L bovine serum albumin solution, the solvent is phosphate buffer solution (pH=7.4), wherein the volume _{of Fe2O3} particle water dispersion, bovine serum albumin solution and zinc chloride aqueous solution The ratio is 10-25:10-25:1. Under mechanical stirring, after reacting at room temperature for 2-8 hours, the bovine serum albumin/Zn ₃ (PO ₄ ) ₂ /Fe ₂ O ₃ hybrid magnetic nanoflowers are obtained through magnetic field separation, water washing and freeze-drying.

Example 3: Preparation of papain/Zn ₃ (PO ₄ ) ₂ /Fe ₃ O ₄ hybrid magnetic nanoflowers

Disperse Fe ₃ O ₄ particles evenly in water to obtain an aqueous dispersion of Fe ₃ O ₄ particles with a mass fraction of 0.05-0.15%, and add it to a three-necked flask with mechanical stirring; prepare nitric acid with a concentration of 0.2-0.5 mol/L Add the zinc aqueous solution into the three-necked flask, and stir it mechanically for 0.5-3h. Then add _0.2-1g /L papain solution, the solvent is phosphate buffer solution (pH=7.2), wherein the volume ratio of _Fe3O4 particle water dispersion, papain solution and zinc nitrate aqueous solution is 10-25 :10～25:1. Under mechanical stirring, after reacting at room temperature for 2-8 hours, the papain/Zn ₃ (PO ₄ ) ₂ /Fe ₃ O ₄ hybrid magnetic nanoflowers are obtained through magnetic field separation, water washing and freeze-drying.

Example 4: Preparation of papain/Zn ₃ (PO ₄ ) ₂ /Fe ₂ O ₃ hybrid magnetic nanoflowers

Disperse Fe ₂ O ₃ particles evenly in water to obtain an aqueous dispersion of Fe ₂ O ₃ particles with a mass fraction of 0.05-0.15%, and add it to a three-necked flask with mechanical stirring; prepare sulfuric acid with a concentration of 0.2-0.5 mol/L Add the zinc aqueous solution into the three-necked flask, and stir it mechanically for 0.5-3h. Then add 0.2-1g/L papain solution, the solvent is phosphate buffer solution (pH=7.4), wherein the volume ratio _{of Fe2O3} particle water dispersion, papain solution and zinc sulfate aqueous solution is 10-25 :10～25:1. Under mechanical stirring, after reacting at room temperature for 2-8 hours, the papain/Zn ₃ (PO ₄ ) ₂ /Fe ₂ O ₃ hybrid magnetic nanoflowers are obtained through magnetic field separation, water washing and freeze-drying.

Example 5: Preparation of trypsin/Zn ₃ (PO ₄ ) ₂ /Fe ₃ O ₄ hybrid magnetic nanoflowers

Disperse Fe ₃ O ₄ particles evenly in water to obtain a water dispersion of Fe ₃ O ₄ particles with a mass fraction of 0.05-0.15%, and add it to a three-necked flask with mechanical stirring; prepare acetic acid with a concentration of 0.2-0.5mol/L Add the zinc aqueous solution into the three-necked flask, and stir it mechanically for 0.5-3h. Then add _0.2-1g /L trypsin solution, the solvent is phosphate buffer solution (pH=7.2), wherein the volume ratio of _Fe3O4 particle water dispersion, trypsin solution and zinc acetate aqueous solution is 10-25 :10～25:1. Under mechanical stirring, after reacting at room temperature for 2-8 hours, magnetic field separation, water washing and freeze-drying are performed to obtain trypsin/Zn ₃ (PO ₄ ) ₂ /Fe ₃ O ₄ hybrid magnetic nanoflowers.

Example 6: Preparation of trypsin/Zn ₃ (PO ₄ ) ₂ /Fe ₂ O ₃ hybrid magnetic nanoflowers

Disperse Fe ₂ O ₃ particles evenly in water to obtain an aqueous dispersion of Fe ₂ O ₃ particles with a mass fraction of 0.05-0.15%, and add it to a three-necked flask with mechanical stirring; prepare nitric acid with a concentration of 0.2-0.5 mol/L Add the zinc aqueous solution into the three-necked flask, and stir it mechanically for 0.5-3h. Then add 0.2-1g/L trypsin solution, the solvent is phosphate buffer solution (pH=7.2), wherein the volume ratio _{of Fe2O3} particle water dispersion, trypsin solution and zinc nitrate aqueous solution is 10-25 :10～25:1. Under mechanical stirring, after reacting at room temperature for 2-8 hours, magnetic field separation, water washing and freeze-drying are performed to obtain trypsin/Zn ₃ (PO ₄ ) ₂ /Fe ₂ O ₃ hybrid magnetic nanoflowers.

Example 7: Preparation of amylase/Zn ₃ (PO ₄ ) ₂ /Fe ₃ O ₄ hybrid magnetic nanoflowers

Evenly disperse Fe ₃ O ₄ particles in water to obtain a water dispersion of Fe ₃ O ₄ particles with a mass fraction of 0.05-0.15%, and add it to a three-necked flask with mechanical stirring; prepare chlorine with a concentration of 0.2-0.5mol/L Zinc chloride aqueous solution was added into a three-necked flask, and mechanically stirred for 0.5-3 hours. Then add 0.2～1g/L amylase solution, the solvent is phosphate buffer solution (pH=7.2), wherein the volume ratio of _Fe3O4 particle water dispersion, amylase solution and zinc chloride aqueous solution is ₁₀ ～ 25:10～25:1. Under mechanical stirring, after reacting at room temperature for 2-8 hours, the amylase/Zn ₃ (PO ₄ ) ₂ /Fe ₃ O ₄ hybrid magnetic nanoflowers are obtained through magnetic field separation, water washing and freeze-drying.

Example 8: Preparation of amylase/Zn ₃ (PO ₄ ) ₂ /Fe ₂ O ₃ hybrid magnetic nanoflowers

Disperse the Fe ₂ O ₃ particles evenly in water to obtain an aqueous dispersion of Fe ₂ O ₃ particles with a mass fraction of 0.05-0.15%, and add it to a three-necked flask with mechanical stirring; prepare acetic acid with a concentration of 0.2-0.5 mol/L Add the zinc aqueous solution into the three-necked flask, and stir it mechanically for 0.5-3h. Then add 0.2-1g/L amylase solution, the solvent is phosphate buffer solution (pH=7.4), wherein the volume ratio _{of Fe2O3} particle water dispersion, amylase solution and zinc acetate aqueous solution is 10-25 :10～25:1. Under mechanical stirring, after reacting at room temperature for 2-8 hours, the amylase/Zn ₃ (PO ₄ ) ₂ /Fe ₂ O ₃ hybrid magnetic nanoflowers are obtained through magnetic field separation, water washing and freeze-drying.

Example 9: Preparation of lipase/Zn ₃ (PO ₄ ) ₂ /Fe ₃ O ₄ hybrid magnetic nanoflowers

Disperse Fe ₃ O ₄ particles evenly in water to obtain a water dispersion of Fe ₃ O ₄ particles with a mass fraction of 0.05-0.15%, and add it to a three-necked flask with mechanical stirring; prepare acetic acid with a concentration of 0.2-0.5mol/L Add the zinc aqueous solution into the three-necked flask, and stir it mechanically for 0.5-3h. Then add _0.2-1g /L lipase solution, the solvent is phosphate buffer solution (pH=7.4), wherein the volume ratio of _Fe3O4 particle water dispersion, lipase solution and zinc acetate aqueous solution is 10-25 :10～25:1. Under mechanical stirring, after reacting at room temperature for 2-8 hours, the lipase/Zn ₃ (PO ₄ ) ₂ /Fe ₃ O ₄ hybrid magnetic nanoflowers are obtained through magnetic field separation, water washing and freeze-drying.

Example 10: Preparation of lipase/Zn ₃ (PO ₄ ) ₂ /Fe ₂ O ₃ hybrid magnetic nanoflowers

Disperse Fe ₂ O ₃ particles evenly in water to obtain an aqueous dispersion of Fe ₂ O ₃ particles with a mass fraction of 0.05-0.15%, and add it to a three-necked flask with mechanical stirring; prepare nitric acid with a concentration of 0.2-0.5 mol/L Add the zinc aqueous solution into the three-necked flask, and stir it mechanically for 0.5-3h. Then add 0.2-1g/L lipase solution, the solvent is phosphate buffer solution (pH=7.2), wherein the volume ratio _{of Fe2O3} particle water dispersion, lipase solution and zinc nitrate aqueous solution is 10-25 :10～25:1. Under mechanical stirring, after reacting at room temperature for 2-8 hours, the lipase/Zn ₃ (PO ₄ ) ₂ /Fe ₂ O ₃ hybrid magnetic nanoflowers are obtained through magnetic field separation, water washing and freeze-drying.

Claims (4)

1. a preparation method for hybrid magnetic Nano flower, is characterized in that step is as follows:

Step 1: magnetic-particle is dispersed in water, being mixed with mass fraction is 0.05 ~ 0.15% magnetic-particle aqueous dispersions, joins with in churned mechanically container;

Step 2: add the soluble Zn saline solution that concentration is 0.2 ~ 0.5mol/L, stirs 0.5-3h; Wherein the volume ratio of soluble Zn saline solution and magnetic-particle aqueous dispersions is 1:10 ~ 25;

Step 3: the PBS adding 0.2 ~ 1g/L, wherein the volume ratio of PBS and soluble Zn saline solution is 10 ~ 25:1; The pH value range of described PBS is 7.2 ~ 7.4; Solute in described PBS is solubility large biological molecule;

Step 4: under mechanical agitation, after room temperature reaction 2 ~ 8h, obtains hybrid magnetic Nano flower through magnetic field separation, washing, freeze drying.

2. the preparation method of hybrid magnetic Nano flower according to claim 1, is characterized in that: described soluble zinc salt is: one or more any than mixing in zinc acetate and hydrate, zinc nitrate and hydrate thereof, zinc chloride and hydrate thereof or zinc sulfate and hydrate thereof.

3. the preparation method that hybrid magnetic Nano is colored according to claim 1, is characterized in that: described solubility large biological molecule is: bovine serum albumin(BSA), pepsin, trypsase, lipase, amylase or papain.

4. the preparation method that hybrid magnetic Nano is colored according to claim 1, it is characterized in that: described PBS is: sodium dihydrogen phosphate, sodium hydrogen phosphate and water, or potassium dihydrogen phosphate, dipotassium hydrogen phosphate and water, or sodium dihydrogen phosphate, dipotassium hydrogen phosphate and water, or potassium dihydrogen phosphate, sodium hydrogen phosphate and water.